Application of direct electrometry in studies of microbial rhodopsins reconstituted in proteoliposomes

Siletsky, Sergey A.; Mamedov, Mahir D.; Лукашев, Е. П.; Balashov, Sergei P.; Petrovskaya, L. E.

doi:10.1007/s12551-022-00986-y

Cited by 6 publications

(1 citation statement)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The data from time resolved optical and photoelectric measurements were fit into a sum of individual exponents using Pluk [ 59 ], Origin (OriginLab Corporation, Northampton, MA, USA), and MATLAB (The Mathworks, South Natick, MA, USA) software, as described earlier for the wild-type and mutants of ESR, BR, cytochrome oxidase, and other proteins [ 43 , 60 , 61 , 62 , 63 , 64 , 65 , 66 ].…”

Section: Methodsmentioning

confidence: 99%

Oriented Insertion of ESR-Containing Hybrid Proteins in Proteoliposomes

Petrovskaya

Лукашев

Mamedov

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

Microbial rhodopsins comprise a diverse family of retinal-containing membrane proteins that convert absorbed light energy to transmembrane ion transport or sensory signals. Incorporation of these proteins in proteoliposomes allows their properties to be studied in a native-like environment; however, unidirectional protein orientation in the artificial membranes is rarely observed. We aimed to obtain proteoliposomes with unidirectional orientation using a proton-pumping retinal protein from Exiguobacterium sibiricum, ESR, as a model. Three ESR hybrids with soluble protein domains (mCherry or thioredoxin at the C-terminus and Caf1M chaperone at the N-terminus) were obtained and characterized. The photocycle of the hybrid proteins incorporated in proteoliposomes demonstrated a higher pKa of the M state accumulation compared to that of the wild-type ESR. Large negative electrogenic phases and an increase in the relative amplitude of kinetic components in the microsecond time range in the kinetics of membrane potential generation of ESR-Cherry and ESR-Trx indicate a decrease in the efficiency of transmembrane proton transport. On the contrary, Caf-ESR demonstrates a native-like kinetics of membrane potential generation and the corresponding electrogenic stages. Our experiments show that the hybrid with Caf1M promotes the unidirectional orientation of ESR in proteoliposomes.

show abstract

Section: Methodsmentioning

confidence: 99%

Oriented Insertion of ESR-Containing Hybrid Proteins in Proteoliposomes

Petrovskaya

Лукашев

Mamedov

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

Editorial for the special issue of biophysical reviews on the 9th Congress of the Russian society for photobiology held in Shepsi, Krasnodar region, Russia, on September 12–19, 2021

et al. 2022

View full text Add to dashboard Cite

Features of the Mechanism of Proton Transport in ESR, Retinal Protein from Exiguobacterium sibiricum

Petrovskaya,

Siletsky,

Mamedov

et al. 2023

Biochemistry Moscow

View full text Add to dashboard Cite

Retinal-containing light-sensitive proteins – rhodopsins – are found in many microorganisms. Interest in them is largely explained by their role in light energy storage and photoregulation in microorganisms, as well as the prospects for their use in optogenetics to control neuronal activity, including treatment of various diseases. One of the representatives of microbial rhodopsins is ESR, the retinal protein of Exiguobacterium sibiricum. What distinguishes ESR from homologous proteins is the presence of a lysine residue (Lys96) as a proton donor for the Schiff base. This feature, along with the hydrogen bond of the proton acceptor Asp85 with the His57 residue, determines functional characteristics of ESR as a proton pump. This review examines the results of ESR studies conducted using various methods, including direct electrometry. Comparison of the obtained data with the results of structural studies and with other retinal proteins allows us to draw conclusions about the mechanisms of transport of hydrogen ions in ESR and similar retinal proteins.

show abstract

Application of direct electrometry in studies of microbial rhodopsins reconstituted in proteoliposomes

Cited by 6 publications

References 47 publications

Oriented Insertion of ESR-Containing Hybrid Proteins in Proteoliposomes

Oriented Insertion of ESR-Containing Hybrid Proteins in Proteoliposomes

Editorial for the special issue of biophysical reviews on the 9th Congress of the Russian society for photobiology held in Shepsi, Krasnodar region, Russia, on September 12–19, 2021

Features of the Mechanism of Proton Transport in ESR, Retinal Protein from Exiguobacterium sibiricum

Contact Info

Product

Resources

About